Difference between revisions of "Part:BBa K4451008"
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BBa_K4451008 encodes the Alc protein from T4 phage. This protein has been reported to interact with the RNA polymerase beta prime subunit and cause host transcription shutoff (Kashlev et al., 1993).
 
BBa_K4451008 encodes the Alc protein from T4 phage. This protein has been reported to interact with the RNA polymerase beta prime subunit and cause host transcription shutoff (Kashlev et al., 1993).
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===Description===
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iGEM Sheffield 2023- PARSE
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A transcriptional terminator from T4 bacteriophage that has been shown to suppress growth rate in E. coli.
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The Alc gene encodes a site-specific termination factor, which binds to commonly found sites in the E. coli genome and causes early transcriptional termination, thus reducing transcriptional output and reducing growth rate. Termination is only possible on actively processing RNA polymerases, any stall within 15bp of the Alc site abolishes early termination. This, alongside AsiA (BBa_K4451010), is utilised to trigger transcriptional redirection toward phage genes in the T4 infection cycle (Kashlev et al., 1993).
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https://static.igem.wiki/teams/4939/wiki/assets/assets/registry-assets/bba-k4451008-meanlog2od.png
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Figure 1: Mean log2(OD) against time for Alc
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From Figure 1, ALc shows a slightly lower mean log2OD compare to vsfGFP after around 7.5 hours. The different [IPTG]/different levels of induction of Alc doesn't show any significant difference in the mean log2OD. This data is inconclusive as to whether Alc protein expression slows the growth of E. coli in comparison to the expression of vsfGFP.
  
 
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Latest revision as of 15:30, 12 October 2023


T4 Alc

BBa_K4451008 encodes the Alc protein from T4 phage. This protein has been reported to interact with the RNA polymerase beta prime subunit and cause host transcription shutoff (Kashlev et al., 1993).

Description

iGEM Sheffield 2023- PARSE

A transcriptional terminator from T4 bacteriophage that has been shown to suppress growth rate in E. coli. The Alc gene encodes a site-specific termination factor, which binds to commonly found sites in the E. coli genome and causes early transcriptional termination, thus reducing transcriptional output and reducing growth rate. Termination is only possible on actively processing RNA polymerases, any stall within 15bp of the Alc site abolishes early termination. This, alongside AsiA (BBa_K4451010), is utilised to trigger transcriptional redirection toward phage genes in the T4 infection cycle (Kashlev et al., 1993).

bba-k4451008-meanlog2od.png

Figure 1: Mean log2(OD) against time for Alc

From Figure 1, ALc shows a slightly lower mean log2OD compare to vsfGFP after around 7.5 hours. The different [IPTG]/different levels of induction of Alc doesn't show any significant difference in the mean log2OD. This data is inconclusive as to whether Alc protein expression slows the growth of E. coli in comparison to the expression of vsfGFP.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 7
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 7
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 413
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 7
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 7
  • 1000
    COMPATIBLE WITH RFC[1000]